Validation of scale‐age determination in European grayling Thymallus thymallus using tag‐recapture analysis

To validate age determination from scales in European grayling Thymallus thymallus, the scale‐read age of fish was compared with the true age obtained by tag‐recapture analysis. A total of 3997 individuals were tagged with visible implant tags and passive integrated transponder (PIT) tags in the River Wylye, south‐west England during 1999–2007. Annual repeat surveys were undertaken and collected scales read without prior knowledge of tag‐recapture age. Accuracy of fish ageing by scales was highest in 1 and 2 year‐old fish but decreased in older fish. In later life stages (>4 years old), underestimation of age occurred and the error in reading scales rose to 51·9% in 5 year‐old fish. Age assigned from scales underestimated the tag‐recapture assigned age by as much as 3 years. This study suggests that use of scales is an appropriate method to age a short‐lived population of T. thymallus inhabiting productive lotic systems. The underestimation of age in older fish, however, needs to be considered in the management of fish stocks because it may lead to undesirable exploitation of population.     

The growth ofOreochromis andersonii (Pisces: Cichlidae) from the Okavango Delta,Botswana, and a comparison of the scale and otolith methods of ageing

Synopsis Otoliths and scales were used for age and growth determination ofOreochromis andersonii from the Okavango Delta, Botswana. Marginal increment analysis showed that an annulus was formed in both the scales and otoliths during the dry summer period. Using scales, the growth ofO. andersonii was described by L_t = 285.27(1-e^{-0.26(t+2.02)}) mm SL and using otoliths by the equation L_t = 267.48(1-e^{-0.25(t+2.18)}) mm SL. Maximum age estimates of 10 years using scales and 13 years using otoliths were obtained and the growth curves were significantly different (p < 0.01). Age estimation using scales tended to over-emphasise growth inO. andersonii resulting in larger predicted lengths-at-age. For this reason, otoliths are considered to be more reliable and suitable than scales in determining the age and growth of this species.     

Age determination in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>: history,relative utility of ageing structures,precision and accuracy

The common carp Cyprinus carpio is one of the most widely-distributed freshwater fishes in the world. Due to its value for conservation and fisheries in several native/translocated areas of distribution and its detrimental effects on the aquatic ecosystem in most invasive areas, robust age-based population dynamics models are required for successful management of this species. The present study provides a global review of age determination in carp, including a historical account of ageing methods, an assessment of the relative utility of ageing structures, and an evaluation of precision and accuracy (i.e. validation) of age estimates. Historically, scales were by far the most widely-employed structure, followed by the operculum, otolith, dorsal spine, vertebra and fin ray. However, in countries where carp is categorised as ‘high risk’ of impact, use of alternative structures to the scale was predominant. Causal criteria analysis showed scales and opercula to provide inconsistent evidence for successful annulus identification/counting, whereas consistent evidence was found for otoliths, dorsal spines, vertebrae and (pectoral) fin rays. Precision was always above reference thresholds for scales, whereas for otoliths, dorsal spines and fin rays was in several cases below. Accuracy was addressed sporadically and mostly in high-risk countries. It is suggested that dorsal spines or pectoral fin rays should be used in lieu of scales as non-lethal ageing structures, and otoliths (or vertebrae, pending more research) otherwise, and that validation should always be attempted as part of the set-up of more appropriate ageing protocols and use of correct terminology.     

Age determination of Anisotremus interruptus (Perciformes: Haemulidae) by scale reading, in the coast of Colima, Mexico

This paper deals with the age determination in Anisotremus interruptus by reading of scales that allowed the identification of 9 age groups. The width of the growth rings diminishes as the age increases. The growth of the scales is proportional to the fish growth. The period of highest growth rate happens during the first year of life. During the first year A. interruptus grows 12.52 cm, the second year 4.95 cm and the third, 4.60 cm. The strategy of the quick growth during the first year of life allows A. interruptus to diminish the natural mortality.     

Comparison of scale and otolith age readings for trahira,Hoplias malabaricus (Bloch, 1794), from Paraná River,Argentina

The aim of this work was to compare age determinations and precision using two deposition structures of trahira Hoplias malabaricus (Bloch, 1794): the scales, which are most frequently used, and otoliths (lapilli). The length‐age relationships were obtained with both structures and compared with results from previous studies. The 163 sets of trahira otoliths (lapilli) and scales were 17–46 cm standard length (SL) from Cayastá (Santa Fe) and Islas Lechiguanas (Entre Ríos), Paraná River, Argentina. Three independent readings of each structure were conducted. An age bias plot was performed to compare age estimations from scales and otoliths. To assess the precision of age determinations using both structures, the percent agreement among readers for both structures and the coefficient of variation were calculated (%CV). The age‐length relationships were plotted and fitted with the von Bertalanffy growth function for both structures and compared with previous works. Age readings recorded for scales were lower than those recorded for otoliths for ages above or equal to 3 years. Percent agreement among readers was higher than 80% for otoliths and less than 65% for scales. The%CV obtained for scales was 20% for young fish and 17% for adults. For otoliths the %CV was 7% for young fish and 3% for adults. The %CV obtained for scales was over the recommended limit (>7.6%). The von Bertalanffy parameters for scales were L_inf = 45.80 mm; k = 0.29; t₀ = ?1.34 and for otoliths were L_inf = 40.76 mm; k = 0.39; t₀ = ?1.05. Precision of age estimations assessed from the percent agreement and the coefficient of variation indicates that the scales of the trahira are inappropriate to estimate age in population studies for juvenile and adult specimens.     

Precision of the age–length increments of three cyprinids: effects of fish number and sub‐sampling strategy

The effects of number of fish that are aged and scale sub‐sampling strategies on the precision of estimates of mean age–length increments from populations of Rutilus rutilus, Leuciscus leuciscus and Leuciscus cephalus were tested. Analyses used data derived from river fish communities in eastern England, U.K.. Regarding the number of fishes analysed in each age group, for each species and mean fork‐length increment at age, significant relationships were detected between sample size (n) and the coefficient of variation of the mean (Z) and mean length increment and measured variance (s²). This enabled calculation of the number of scales for producing a mean length increment at age according to . Outputs indicated that the number of scales requiring ageing increased substantially as precision increased, but with little variation between species per age category. Ageing between seven and 12 scales per age group would thus provide estimates at 10% precision. As the ages of fishes are not known in advance of scale ageing, the effect of scale sub‐sampling regime on precision was also tested using randomized strategies of 10 fish per 5 mm, five per 5 mm, three per 5 mm, 10 per 10 mm, five per 10 mm and three per 10 mm. These were applied to the datasets and the consequences of their reduction in the number of scales for precision were determined using . When compared to no sub‐sampling, three per 10 mm always significantly reduced data precision, whereas 10 per 5 mm never significantly reduced precision. These outputs can thus be applied to the design of fish sampling protocols where age and growth estimates are required, with the randomized sub‐sampling likely to be the most useful strategy.     

Life-history strategies of Acrossocheilus fasciatus (Barbinae, Cyprinidae) in the Huishui Stream of the Qingyi watershed, China

Life-history traits of Acrossocheilus fasciatus were examined using 384 specimens collected monthly during May 2009 and April 2010 in the Huishui Stream of the Qingyi watershed, China. Using scales for age determination, female and male fish comprised five and four age groups, respectively. The monthly changes in marginal increment ratio suggested that annuli on scales were formed during March through May. Total lengths back-calculated significantly increased with age for both sexes and varied significantly between the two sexes at each age. The fact that females had larger body size and grew faster than males indicated the sexual size dimorphism for this species. Both sexes got their 50% maturity at age 3, when females and males were 105.3 and 112.1?mm total length, respectively. Based on the monthly changes in the gonado-somatic index and egg-development process, fish spawned from April through August. Absolute fecundity ranged from 295 to 3,573 eggs per fish and increased significantly with age. But relative fecundity, ranging from 11.77 to 69.96?eggs/g, was not significantly different among age groups. Compared with the life-history traits of an upstream population in the Puxi Stream (a headwater stream within this study watershed), the downstream population of A. fasciatus in the Huishui Stream (a 4th-order stream) exhibits larger body size, faster somatic growth, later sexual maturity, and lower reproductive investment. These variations in life-history strategies between the two populations could perhaps be explained by the spatial heterogeneity in habitat environment along the upstream–downstream gradient in this watershed.     

The growth of the freshwater murrel,Ophicephalus punctatus Bloch

Summary The growth of O. punctatus has been studied from the zones on the opercular bones and scales. Size estimates obtained from the opercular bones and scales were further substantiated by length frequency distribution and back-calculations. A close agreement in length-age relationship was obtained by various methods. These observations provide adequate evidence towards the validity of age determination in O. punctatus.Growth rate differs markedly in the two sexes. Males grow faster than the females. To study the changes in length with age, von Bertalanffy growth equation and Gompertz curve were used. The theoretical lengths obtained from the von Bertalanffy equation agreed very closely with the observed lengths.There is sexual difference in the weight-length relationship of O. punctatus. Modal weight of each year class obtained according to age reading showed that growth in weight is faster in males. The theoretical growth equation gives a good fit for weight-age data. O. punctatus is generally a fish of the impounded waters. The interrelationship between pond environment and growth characteristics has been discussed.     

Determination of relative age using growth increments of scales as a minimally invasive method in the tropical freshwater Apteronotus leptorhynchus

This study describes a method for the determination of relative age in a tropical teleost, the brown ghost knifefish Apteronotus leptorhynchus. This method is based on identification of the maximum number of scale circuli, which is thought to be associated with the oldest scales, and thus to be the most indicative of the age of a given fish. Relative age can be inferred by relating differences in maximum circulus counts to the average rate of circulus addition, which was estimated at 34 circuli per year in adult fish through oxytetracycline marking. This method shows high inter‐investigator reliability and has a limited effect on fish because of the low number of scales required in order to determine the maximum number of circuli with a sufficiently high confidence level. Analysis of the frequency distribution of the circulus counts revealed periodic patterns that are similar among fish, presumably reflecting the environmental life history of the individuals. Regression analysis and comparison of addition rates showed that scale circulus counts and otolith ring counts are equivalent approaches for age estimation, but scale analysis is superior because of its limited invasiveness and the lower demand in terms of technical skills and expensive instrumentation.     

Age determination and growth of the pollan, Coregonus autumnalis pollan Thompson, of Lough Neagh, Northern Ireland

Scales from Lough Neagh pollan display a large number of checks, making age determination difficult. Sclerite counts showed that an annual check is formed on scales in May and a second accessory check in most young fish in October. The method of ageing from scales was supported by inspection of length-frequency plots and by following the growth of pollan in their first 2 years of life. The body-scale relationship was curvilinear. Back-calculation showed that pollan of both sexes attain a fork length of 29 cm in 5 years (1 ∞=28.9 cm; k = 0.65; l ₀= -0.06 year). There is no evidence that annual growth rates have changed since 1965. Possible environmental causes of scale check formation are discussed.     

Age determination in the Red fox (Vulpes vulpes)–an evaluation of technique efficiency as applied to a sample of suburban foxes

A review of age determination techniques that have been applied to the family Canidae is presented. It is shown that the pattern of growth and development of the various species is very similar, the rate of development being greater in the smaller, shorter-lived species, and vice versa.
The occurrence of annuli in the hard tissues of London foxes is demonstrated; these annuli can be used reliably for age determination, despite suggestions that temperate zone animals from weakly continental climates have indistinct annuli. The objective ages determined by cementum annuli are then used to investigate the value of less time-consuming age determination techniques, particularly in the separation of the young-of-the-year from adults. The results obtained from different studies vary, and these differences are discussed. It is suggested that the speed of development varies slightly in different Red fox populations, and so caution must be exercised before data from one population areapplied to another population.     

Age validation of juvenile Notothenia rossii at Potter Cove, South Shetland Islands, using mark-recapture data

Among all validation methods of age determination in fish, release of known age and marked specimens gives the most reliable information. We carried out a tag-recapture experiment on Notothenia rossii at Potter Cove, to validate, for first time for this species using this method, the principle of annual deposition of an annulus in scales and otoliths. Of 132 juvenile specimens (TL = 22.1–38.1 cm) tagged and released in successive years from 2004 to 2010, 7 were recaptured at the same site after periods of 1–13 months. In scales of five specimens recovered after 10–13 months, one extra annulus was laid down, exhibiting an additional winter zone of closely spaced sclerites. Consistently, the same analysis in two individuals marked and recaptured during the same summer, after 1–3 months at liberty, did not show the deposition of an additional annulus. All the fish tagged or recaptured during the experiment period (December to March) showed in their scales an edge zone of widely spaced sclerites, in agreement with the known pattern of growth in summer. Likewise, an analysis in selected specimens showed good consistency between the numbers of sclerites deposited in scales and the time of fish release. The comparative analysis between scales taken at recapture and otoliths of the same individual allowed a simultaneous counting of the annuli with complete correspondence. The growth in length of fish ranged from 0.5 to 6.1 cm, depending on the time of release.     

<Emphasis Type="Italic">Strobilomyces verruculosus</Emphasis> sp. nov. from Japan

A new species, Strobilomyces verruculosus, is described and illustrated. It is morphologically distinct from other species of Strobilomyces by having a verruculose pileus with small subpyramidal scales, a long and thick stipe with small warty to appressed scales, subdecurrent tubes, and incompletely reticulate basidiospores. Molecular phylogenetic analyses using the mitochondrial cytochrome oxidase subunit 3 (cox3) gene support that it belongs to the genus Strobilomyces and is highly differentiated from the other members of this genus found in Japan.     

Two New Species of Goby of the Genus <Emphasis Type="Italic">Astrabe</Emphasis> from Japan

Two species of goby belonging to the genus Astrabe are described from Japan as new species, A. flavimaculata and A. fasciata. A. flavimaculata is distinguishable from A. lactisella, the type species and hitherto the only known species of the genus, in that it has no protrusion on the upper posterior part of the dermal fold along the upper margin of the eye, fewer scales in a longitudinal row, predorsal scales, scales on the belly, a narrower white transverse band across the base of the pectoral fins, and in life yellow markings on a dark brown ground colour except for the white transverse band across the base of the pectoral fins. A. fasciata is distinguishable from A. lactisella in that it has fewer scales in a transverse row, a narrower scaled area on the lateral side of the body, a narrower white transverse band across the base of the pectoral fins, and a white transverse band across the anterior part of the 1st dorsal fin extending to the ventral side of the body.     

Automatic Nuclei Segmentation in H&E Stained Breast Cancer Histopathology Images

The introduction of fast digital slide scanners that provide whole slide images has led to a revival of interest in image analysis applications in pathology. Segmentation of cells and nuclei is an important first step towards automatic analysis of digitized microscopy images. We therefore developed an automated nuclei segmentation method that works with hematoxylin and eosin (H&E) stained breast cancer histopathology images, which represent regions of whole digital slides. The procedure can be divided into four main steps: 1) pre-processing with color unmixing and morphological operators, 2) marker-controlled watershed segmentation at multiple scales and with different markers, 3) post-processing for rejection of false regions and 4) merging of the results from multiple scales. The procedure was developed on a set of 21 breast cancer cases (subset A) and tested on a separate validation set of 18 cases (subset B). The evaluation was done in terms of both detection accuracy (sensitivity and positive predictive value) and segmentation accuracy (Dice coefficient). The mean estimated sensitivity for subset A was 0.875 (±0.092) and for subset B 0.853 (±0.077). The mean estimated positive predictive value was 0.904 (±0.075) and 0.886 (±0.069) for subsets A and B, respectively. For both subsets, the distribution of the Dice coefficients had a high peak around 0.9, with the vast majority of segmentations having values larger than 0.8.     

A comparison of back-calculated lengths of silver carp derived from bony structures

Linear relationships were found between body length–scale radius, body length-cleithrum length and body length-urohyal length ( r =0·949, 0·984 and 0·974 respectively) in silver carp Hypophthalmichthys molitrix. Out of 180 comparisons, which were made between different methods of back-calculating length and ageing structures at ages 2-7 years, the differences were not significant ( P ≥ 0·05) in 132 (73·3%) cases. There were no significant differences among different methods in older age classes and between the Fraser-Lee method and body proportional method (BPH) at all ages. The results demonstrated that in all structures, back-calculated lengths estimated by the scale proportional hypotheses (SPH) were less than the other methods. For scales, SPH was similar to the direct proportional method of Dahl-Lea (DPM) but for cleithra and urohyal bones it was similar to BPH and Fraser-Lee. As the fish increased in age, the differences between back-calculated lengths decreased. When lengths were back-calculated using cleithra and urohyal bones, differences between the structures at age 2 years were 17 mm from DPM, 33 mm from SPH, 37 mm from BPH and 37 mm from Fraser-Lee, while at age 8 years mean differences were 7 mm from DPM and 8 mm from the SPH, BPH and Fraser-Lee methods. It is suggested that the differences would be less if: (1) back-calculated lengths are based on a large number of random samples; (2) measurements of scale radius, cleithrum length and urohyal length were precise; (3) determination of focus or origin were precise; (4) the sampling of scales were from the same row.     

Methods of age determination of band-finned flounder <Emphasis Type="Italic">Liopsetta pinnifasciata</Emphasis> (Pleuronectidae)

Techniques involving chemical treatment are suggested for discrimination of annual and seasonal rings in the otoliths of band-finned flounder Liopsetta pinnifasciata. In the first annual zone, three seasonal rings are discerned, and four seasonal rings are discerned in subsequent zones. It is supposed that these rings correspond to biological seasons. The annual rings on otoliths and scales are formed synchronously: the first ring in December–March, and the subsequent ones in December–February. The time of formation of the first annual zone is less than a year; accordingly, the age of Liopsetta pinnifasciata determined by registering structures is less than its calendar age. According to calculations, metamorphosis of Liopsetta pinnifasciata ends at the length approximately 20 mm.     

Mineralized scale patterns on the cell periphery of the chrysophyte Mallomonas determined by comparative 3D Cryo-FIB SEM data processing

Unicellular protists can biomineralize spatially complex and functional shells. A typical cell of the photosynthetic synurophyte Mallomonas is covered by about 60–100 silica scales. Their geometric arrangement, the so-called scale case, mainly depends on the species and on the cell cycle. In this study, the scale case of the synurophyte Mallomonas was preserved in aqueous suspension using high-pressure freezing (HPF). From this specimen, a three-dimensional (3D) data set spanning a volume of about 25.6 μm × 19.2 μm × 4.2 μm with a voxel size of 12.5 nm × 12.5 nm × 25.0 nm was collected by Cryo-FIB SEM in 3 h and 24 min. SEM imaging using In-lens SE detection allowed to clearly differentiate between mineralized, curved scales of less than 0.2 μm thickness and organic cellular ultrastructure or vitrified ice. The three-dimensional spatial orientations and shapes of a minimum set of scales (N = 13) were identified by visual inspection, and manually segmented. Manual and automated segmentation approaches were comparatively applied to one arbitrarily selected reference scale using the differences in grey level between scales and other constituents. Computational automated routines and principal component analysis of the experimentally extracted data created a realistic mathematical model based on the Fibonacci pattern theory. A complete in silico scale case of Mallomonas was reconstructed showing an optimized scale coverage on the cell surface, similarly as it was observed experimentally. The minimum time requirements from harvesting the living cells to the final scale case determination by Cryo-FIB SEM and computational image processing are discussed.     

Age Determination of Bovine Foetuses

The need for methods for age determination of foetuses is defined. X-ray examination of the foetal head is recommended as the surest basis for age determination. Calculations are made concerning the correlation between cranial length and foetal age, and the methods used for measurements are described in detail. Measurements of the metacarpus and metatarsus may also be used, but they give a poorer correlation. A formula is worked out for age determination based on cranial measurements. On x-ray photographs, tooth development is seen to occur in an obvious sequence. This is described. However, the interpretation of these observations is subjective and requires control material for comparison. The same is true for the sequence of ossification of the skeleton, which in foetuses < 120 days gives the surest basis for evaluation. Here, too, a wide range of control material for comparison is necessary. The sequence in the appearance of body hair — development and eruption — is noted, as far as the head is concerned. This alone gives an insecure basis for evaluation. By using the 4 systems of parameters mentioned, the age of a foetus can be determined within an error of ± 4—5 days.     

A new amphipathy scale: I. Determination of the scale from molecular dynamics data

Two new amphipathy scales elaborated from molecular dynamics data are presented. Their applications contribute for the identification of the hydrophobic or hydrophilic regions in proteins solely from the primary structure. The new amphipathy coefficients (AC) reflect the side chain/solvent molecules configurational energies. A polar (water) and an apolar solvent, CCl₄, were used resulting in the two AC_water and AC_CCl4 scales. These solvents were chosen to simulate the aqueous phases and the transmembrane ambients of cellular membranes where the membrane proteins act. The new amphipathy scales were compared with some previous scales determined by different methods, which were also compared between them, indicating more than 90% of the correlation coefficients are less than 0.9: the scales are strictly dependent on the methodologies used in their determination. The AC_CCl4 scale is related with the size of side chain amino acids while AC_water is related with the hydrophobicity of side chain amino acids. The quality of the scales was confirmed by an example of application where AC_water was able to identify correctly the transmembrane, hydrophobic regions of a membrane protein. These results also indicate that water is an important factor responsible for the tertiary structure of membrane proteins.